Fig. 3: N1DARP overexpression inhibits tumorigenesis and stem cell traits of patient-derived organoids and a genetically engineered mouse model. | Cell Discovery

Fig. 3: N1DARP overexpression inhibits tumorigenesis and stem cell traits of patient-derived organoids and a genetically engineered mouse model.

From: A microprotein N1DARP encoded by LINC00261 promotes Notch1 intracellular ___domain (N1ICD) degradation via disrupting USP10-N1ICD interaction to inhibit chemoresistance in Notch1-hyperactivated pancreatic cancer

Fig. 3

a Establishment of pancreatic cancer organoids and cells transfected with vector or N1DARP through subcutaneous injection into nude mice. b Bright-field image and immunohistochemistry analysis of PDAC-1 and PDAC-2 showing their successful establishment. c Left: Microscopic inspection of organoid diameter of PDAC-1 and PDAC-2 transfected with vector or N1DARP. Right: Calculated average organoid area of PDAC-1 and PDAC-2. d Tumor volume of transfected PDAC-1 and PDAC-2 through subcutaneous injection of vector or N1DARP. e Left: Calcein-AM/PI staining visualized by fluorescence microscopy detecting apoptotic cells of PDAC-1 and PDAC-2 organoid with control or N1DARP overexpression followed by treatment with GEM for 72 h. The green signal indicates living tumor organoid, while red dots denote apoptotic cells. Right: quantified apoptotic rate of PDAC-1 and PDAC-2 with vector or N1DARP transfection treated with GEM detected by flow cytometry. f Tumor volume and apoptosis rate detected by flow cytometry of subcutaneously injected mice model using wild-type or N1DARP overexpressed PDAC-1 treated with GEM (40 mg/kg, biweekly). g Generation of KPC and KPNC mice. h Survival analysis with follow-up data of KPC and KPNC. i Tumor weight of KPC and KPNC mice at 16 weeks of age. j Hematoxylin and eosin (HE) staining and immunohistochemistry analysis of Ki-67 and SOX-2 using spontaneous pancreatic tumor from KPC and KPNC using western blot analysis. k Detection of biomarkers of stem-like properties using western blot analysis of spontaneous pancreatic tumor from KPC and KPNC. The data are presented as the mean ± SD of three independent experiments. *P < 0.05, **P < 0.01 by Student’s t-test (c, e, f for apoptosis rate; i for tumor weight); *P < 0.05, **P < 0.01 by one-way ANOVA (d, f for tumor volume); P < 0.05 was considered statistically significant by log-rank test (h). Scale bars, 50 μm (b, j), 200 μm (c, e).

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